The Key Laboratory of Bioactive Materials, Ministry of Education, College of Life Science, Nankai University, Tianjin, 300071, China.
State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300350, China.
Genes Genomics. 2023 May;45(5):681-692. doi: 10.1007/s13258-022-01362-6. Epub 2023 Jan 3.
N-cadherin-mediated cell adhesion is a vital inductor for mesenchymal condensation in chondrogenesis. Recent studies have revealed the involvement of E-cadherin in enhancing the multipotency of mesenchymal stem cells (MSCs) and limb development; however, the signaling crosstalk of E/N-cadherin remains unclear.
This study aimed to explore the synergistic modulation of E/N-cadherin in the chondrogenic differentiation of MSC aggregates.
Human E/N-cadherin-functionalized (hE/N-cad-Fc) poly (lactic-co-glycolic acid) (PLGA) microparticles (hE/N-cad-PLGA) were incorporated into the human MSC (hMSC) aggregates to upregulate the expression of the corresponding endogenous cadherin. The chondrogenic differentiation of the hMSC aggregates was initiated by hE/N-cad-PLGA, controlling the release of transforming growth factor-β (TGF-β). A transcriptome analysis was used to assess differentially expressed genes (DEGs) modulated by hE/N-cad-Fc in hMSC aggregate chondrogenesis. Gene functions and signaling pathways were assessed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. The associated biological pathways were assessed by a protein-protein interaction (PPI) network analysis, and the results were further confirmed by real-time quantitative PCR (qPCR) and western blotting.
A total of 1083 DEGs, comprising 111 upregulated and 972 downregulated genes, were discovered to be related to the enhanced chondrogenic differentiation modulated by hE/N-cad-Fc. The GO and KEGG functional enrichment analyses revealed that hE/N-cad-Fc synergistically regulated the p53-related survival signaling pathway. PPI analysis revealed that mitogen-activated protein kinases (MAPK) caspase regulation is a core aspect of the chondrogenic differentiation process, confirmed by western blotting.
To the best of our knowledge, our study is the first to reveal that the synergistic modulation of E/N-cadherin enhances the chondrogenic differentiation of hMSCs via the ERK1/2-p53 signaling axis.
N-钙黏蛋白介导的细胞黏附是软骨形成中间充质凝聚的重要诱导因子。最近的研究表明 E-钙黏蛋白参与增强间充质干细胞（MSCs）的多能性和肢体发育；然而，E/N-钙黏蛋白的信号串扰尚不清楚。
本研究旨在探讨 E/N-钙黏蛋白在 MSC 聚集物的软骨分化中的协同调节作用。
将人 E/N-钙黏蛋白功能化（hE/N-cad-Fc）聚乳酸-羟基乙酸共聚物（PLGA）微球（hE/N-cad-PLGA）掺入人 MSC（hMSC）聚集物中，以上调相应内源性钙黏蛋白的表达。hE/N-cad-PLGA 启动 hMSC 聚集物的软骨分化，控制转化生长因子-β（TGF-β）的释放。通过转录组分析评估 hE/N-cad-Fc 在 hMSC 聚集物软骨形成中调节的差异表达基因（DEGs）。使用基因本体论（GO）和京都基因与基因组百科全书（KEGG）富集分析评估基因功能和信号通路。通过蛋白质-蛋白质相互作用（PPI）网络分析评估相关生物途径，并通过实时定量 PCR（qPCR）和蛋白质印迹进一步验证。
共发现 1083 个 DEGs，包括 111 个上调基因和 972 个下调基因，这些基因与 hE/N-cad-Fc 增强的软骨分化有关。GO 和 KEGG 功能富集分析表明，hE/N-cad-Fc 协同调节 p53 相关的存活信号通路。PPI 分析表明，丝裂原激活蛋白激酶（MAPK）caspase 调节是软骨分化过程的核心方面，这一点通过蛋白质印迹得到了证实。
据我们所知，我们的研究首次表明，E/N-钙黏蛋白的协同调节通过 ERK1/2-p53 信号轴增强 hMSC 的软骨分化。
